Jie Yao

Intrahepatic mucinous cholangiocarcinoma (IMCC) is a rare subtype of intrahepatic cholangiocarcinoma (IHCC). Limited data describe the genetic characteristics of IMCC and insights on its pathogenesis are lacking. Here, we employed a multi-omics approach to analyze somatic mutations, transcriptome, proteome and metabolome of tumor tissue obtained from a case of IMCC in order to clarify the pathogenesis of IMCC. A total of 54 somatic mutations were detected, including a G12D mutation in KRAS that is likely to be involved in the onset of IMCC. The genes consistently up-regulated at the transcription level and in the proteome were enriched for mucin and mucopolysaccharide biosynthesis, for cell cycle functions and for inflammatory signaling pathways. The consistently down-regulated genes were enriched in bile synthesis and fatty acid metabolism pathways. Further multi-omics analysis found that mucin synthesis by MUC4 and MUC16 was elevated by up-regulated expression of mesothelin (MSLN). Moreover, transcription factor ONECUT3 was identified that possibly activates the transcription of mucin and mucopolysaccharide biosynthesis in IMCC.

Objectives: Multi-drug resistance (MDR) emerged as a serious threat in intensive care unit (ICU) settings. Our study aimed to investigate the major pathogens in ICU and identify the risk factors for MDR infection. Methods: We performed a retrospective analysis of patients admitted to the ICU. Multivariate logistic regression was applied to identify the independent predictors, and then a nomogram to predict the probability of MDR infection. Results: A total of 278 patients with 483 positive cultures were included. 249 (51.55%) had at least one MDR pathogen, including extensively drug-resistant (XDR) 77 (30.92%) and pan drug-resistant (PDR) 39 (15.66%), respectively. Klebsiella pneumonia was the most frequently isolated pathogen. We identified the number of bacteria (OR 2.91, 95% CI 1.97– 4.29, P < 0.001), multiple invasive procedures (OR 2.23, 95% CI 1.37– 3.63, P = 0.001), length of stay (LOS) (OR 1.01, 95% CI 1.00– 1.02, P = 0.007), Hemoglobin (Hb) (OR 0.99, 95% CI 0.98– 1.00, P = 0.01) were independent risk factors for MDR infection. Our nomogram displayed good discrimination with curve AUC was 0.75 (95% CI: 0.70– 0.81). The decision curves also indicate the clinical utility of our nomogram. Additionally, the in-hospital mortality with MDR pathogens was independently associated with XDR (HR, 2.60; 95% CI: 1.08– 6.25; P = 0.03) and total protein (TP) (HR, 0.95; 95% CI: 0.91– 0.99; P = 0.03). Conclusion: The number of bacteria, multiple invasive procedures, LOS, and Hb were the independent predictors associated with MDR pathogens. Our nomogram is potentially useful for predicting the occurrence of MDR infection. Besides, we also identify XDR and TP as the independent risk factors for in-hospital mortality with MDR infection. The current prevalence of MDR strains was also described. The results will contribute to the identification and preventive management of patients at increased risk of infection.

SHARPIN is a tumor‐associated gene involved in the growth and proliferation of many tumor types. A function of SHARPIN in cholangiocarcinoma (CCA) is so far unclear. Here, we studied the role and function of SHARPIN in CCA and revealed its relevant molecular mechanism. The expression of SHARPIN was analyzed in cholangiocarcinoma tissues from patients using immunohistochemistry, quantitative PCR, and western blot analysis. Expression of SHARPIN was suppressed/overexpressed by siRNA silencing or lentiviral overexpression vector, and the effect on cell proliferation was determined by the CCK‐8 assay and flow cytometry. Accumulation of reactive oxygen species was measured with MitoTracker, and JC‐1 staining showed mitochondrial fission/fusion and mitochondrial membrane potential changes as a result of the silencing or overexpression. The ferroptosis marker solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), and the antioxidant enzymes superoxide dismutase 1 (SOD‐1) and SOD‐2 were analyzed by western blot. The results showed that SHARPIN expression was increased in CCA tissue, and this was involved in cell proliferation. SHARPIN silencing resulted in accumulated reactive oxygen species, reduced mitochondrial fission, and a reduced mitochondrial membrane potential. Silencing of SHARPIN inhibited the ubiquitination and degradation of p53, and downregulated levels of SLC7A11, GPX4, SOD‐1, and SOD‐2, all of which contributed to excessive oxidative stress that leads to ferroptosis. Overexpression of SHARPIN would reverse the above process. The collected data suggest that in CCA, SHARPIN‐mediated cell ferroptosis via the p53/SLC7A11/GPX4 signaling pathway is inhibited. Targeting SHARPIN might be a promising approach for the treatment of CCA.

Metabolic reprogramming is a sign of malignant tumors, and targeting the metabolism of tumor cells has become a promising therapeutic approach. Here, we report that Silybin (a nontoxic flavonoid commonly used for liver protection) exhibits prominent anti‐tumor effects on human ovarian cancer cells. Treatment of an ovarian cancer cell line with Silybin interfered with glutamine metabolism and the tricarboxylic acid cycle. We applied the drug affinity responsive target stability approach to show that Silybin binds to isocitrate dehydrogenase 1 (IDH1). This combination leads to reduced phosphorylation of IDH1 and inhibits enzyme activity. IDH1 dysfunction significantly increases the ratio of NADP/NADPH in the cell, causing an increase in reactive oxygen species generation. Immunohistochemistry demonstrated that IDH1 was increased in ovarian cancer samples compared with normal para‐tumoral tissues. Xenograft murine experiments indicated that Silybin administered orally suppressed the growth of the tumor formed by ovarian cancer cells. In combination, our data strongly suggest that Silybin targets IDH1 in ovarian cancer cells and may be a novel treatment candidate.

Lung adenocarcinoma (LUAD) is a disease with high morbidity and mortality rates globally. Holliday junction‑recognizing protein (HJURP) has recently been shown to be a potentially useful biomarker for diagnosing and determining the progression and prognosis of different cancer types. The present study assessed the prognostic value of HJURP expression in LUAD and investigated the biological pathways related to HJURP that are involved in LUAD pathogenesis. It was found that high HJURP expression was significantly associated with stage (P=0.001), T grade (P=0.012) and N grade (P=0.012). Overall survival analysis demonstrated that patients with LUAD and high HJURP expression had a worse prognosis compared with those patients with low HJURP expression (P<0.001). Multivariate analysis using the Cox proportional hazards model indicated that the expression of HJURP [hazard ratio (HR), 1.32; 95% confidence interval (CI), 1.09‑1.60; P=0.004] and stage (HR, 1.90; 95% CI, 1.19‑3.03; P=0.007) were independent prognostic factors for patients with LUAD. Gene set enrichment analysis results showed that genes involved with ‘basal transcription factors’, the ‘cell cycle’, ‘homologous recombination’, ‘non‑small cell lung cancer’ (NSCLC), ‘oocyte meiosis’, ‘p53 signaling pathway’, ‘pathways in cancer’, ‘RNA degradation’ and ‘spliceosome’ were differentially enriched in the high HJURP expression phenotype. Significant correlations were also found between HJURP and several tumor‑infiltrating immune cells, immunomodulators and immune subtypes. Furthermore, western blotting and qPCR analyses confirmed that HJURP was significantly increased in cell lines of NSCLC. In summary, HJURP may be a potentially useful prognostic molecular biomarker of a poor prognosis in LUAD cases. Further experiments are needed to demonstrate the biological effects of HJURP.

Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is associated with high mortality and has become a major public problem threatening patients. This study aimed to explore risk factors for death in patients with Klebsiella pneumoniae (KP) and identify risk factors for CRKP infection. Methods: The study retrospectively analyzed clinical characteristics and microbiological data from patients infected with KP from January 2019 to October 2021 to identify risk factors and mortality, using multivariate logistic regression analysis and Cox regression analysis. Results: A total of 214 KP inpatients were enrolled in our study. The in-hospital mortality rate was significantly higher in patients infected with CRKP (13/68, 19.12%) than carbapenem-susceptible KP (CSKP) (2/146, 1.37%) and the difference was statistically significant (P= 0.03). Multivariate Cox regression analysis showed CRKP isolation (HR 12.26, 95% CI 2.43– 61.68, P = 0.002), lower TP (HR 10.50, 95% CI 1.33– 82.76, P = 0.03), antibiotic days of therapy > 15 (HR 0.08, 95% CI 0.01– 0.56, P= 0.01) and length of stay (LOS) (HR 0.03, 95% CI 0.002– 0.61, P= 0.02) were independent risk factors for death from KP. Additionally, intensive care unit (ICU) stay (OR 21.69, 95% CI 4.50– 118.76, P< 0.001) and previous carbapenem exposure (OR 5.26, 95% CI 1.38– 21.19, P= 0.02) are independent risk factors for CRKP. Conclusion: Our findings showed that patients infected with CRKP have a higher in-hospital mortality rate. Identifying the independent risk factors for CRKP infection may contribute to the management of CRKP and reduce the mortality of KP patients.

Chemokine (C-X-C motif) ligand 1 (CXCL1) is mainly expressed on neutrophils and macrophages and has neutrophil chemoattractant activity. However, natural killer (NK) cells also express CXCL1. We were curious about the role played by CXCL1 in NK cells. Knocking out CXCL1 in hematopoietic cells does not affect the occurrence of NK cells; however, it does hinder NK cell maturity. CXCL1 deletion enhances the expression of immature markers and decreases the expression of functional markers in NK cells, which may explain why it hinders the maturation of NK cells. Specific knockout of CXCL1 in NK cells (CXCL1^flox/flox Ncr1-cre) leads to impaired IFN-γ production and degranulation of NK cells. The lack of CXCL1 may prevent IFN-γ production and degranulation of NK cells by inhibiting the phosphorylation of AKT_S473 and S6. Therefore, we have discovered a new role for CXCL1 in regulating NK cell development and immune surveillance, providing a novel theoretical basis for immunotherapy based on NK cells and potential therapeutic targets for the clinical use of NK cells. Graphical abstract 1. Knockout of CXCL1 in hematopoietic cells inhibits the maturation of NK cells. 2. Knockout of CXCL1 in NK cells inhibits the clearance of lymphoma by NK cells and reduces IFN-γ production and CD107 expression in NK cells. 3. CXCL1 activates the PKD2/mTOR signaling pathway, and promotes the production of IFN-γ and the expression of CD107a in NK cells.

Excitatory neurotoxicity caused by the accumulation of glutamate in the synaptic cleft is an important cause of Parkinson’s disease (PD). Astrocyte glutamate transporter 1 (GLT-1) is the main transporter responsible for transporting glutamate, and investigations toward the regulation of GLT-1 in astrocytes can reveal important insights. Vitamin C (VC) has important protective effects on the brain, but its effect on the regulation of GLT-1 expression is unclear. The purpose of this study was to explore any regulatory effect of VC on GLT-1 expression in astrocytes and to clarify the possible mechanism of such regulation. We found that GLT-1 expression was impaired in 1-methyl-4-phenylpyridinium iodide (MPP⁺)-treated astrocytes, and the transport capacity for glutamate was significantly reduced. Pretreatment with VC restored the GLT-1 expression in the MPP⁺-treated astrocytes. Intraperitoneal VC administration in a PD murine model confirmed that GLT-1 expression was restored in midbrain tissue. The VC-dependent rescue of GLT-1 expression in the MPP⁺-treated astrocytes was shown to be due to inhibition of GLT-1 ubiquitination. Transcriptome sequence analysis revealed a number of differentially expressed genes as a result of VC treatment on MPP⁺-treated astrocytes, including the downregulation of HECT Domain E3 ubiquitin protein ligase 1 (Hectd1). After knocking down Hectd1, the impaired GLT-1 expression caused by MPP⁺ was alleviated, while overexpression of Hectd1 significantly reduced the expression of GLT-1. After overexpression of Hectd1, VC could no longer increase GLT-1 expression of MPP⁺-treated astrocytes, indicating that HECTD1 is essential for VC regulation of GLT-1. Thus, VC reduces the ubiquitination of GLT-1 in astrocytes by inhibiting the expression of HECTD1. Our findings have identified a novel mechanism by which VC regulates the expression of GLT-1 in astrocytes.